Supplementar material for Meer KS, Wheeler JD, Houlihan E, Mullineaux LS (2018) Desperate planktotrophs: decreased settlement selectivit with age in competent eastern oster (Crassostrea virginica) larvae. Mar Ecol Prog Ser Table S1. Average size and percentage of larvae with eespots at each age in the pilot experiment. N, number of individuals sub-sampled. Intervals represent standard deviation. Experiment Hours postcompetenc N Length (μm) Height (μm) % with eespots Pilot 0 22 273 ± 22 262 ± 17 54 4 26 276 ± 16 271 ± 19 54 8 30 287 ± 18 271 ± 25 60 12 30 298 ± 26 287 ± 16 70 Table S2. Analsis of variance in settlement behaviors in response to settlement cue and larval age in the pilot experiment. Two-wa ANOVA tests for differences in the proportion of larvae remaining on the bottom of experimental flasks and larval swimming velocities. Factors are Cue (presence or absence of a settlement cue) and Age (larval age in hours post-competenc. Significant p-values (<0.05) shown in bold. Behavioral metric Experiment Factor F p Proportion of larvae Pilot Cue 40.1 < 0.001 staing on bottom Age 0.47 0.650 Cue x Age 0.53 0.605 Velocit of downward- Pilot Cue 9.15 0.004 swimming larvae Age 1.69 0.189 Cue x Age 2.34 0.092
Table S3. Analsis of variance of explorator behaviors in response to settlement cue and larval age in the pilot experiment. Two-wa ANOVA tests for differences in the upward swimming velocit of larvae, average gross:net distance ratio (GNDR), and proportion swimming in helices. Factors are Cue (presence or absence of a settlement cue) and Age (larval age in hours post-competenc. Significant p-values (<0.05) shown in bold. Behavioral metric Experiment Factor F p Velocit of upward- Pilot Cue 8.59 0.007 swimming larvae Age 0.24 0.868 Cue x Age 6.95 0.001 GNDR of downward- Pilot Cue 12.7 0.001 swimming larvae Age 1.24 0.310 Cue x Age 4.45 0.010 GNDR of upward- Pilot Cue 2.78 0.108 swimming larvae Age 0.200 0.895 Cue x Age 0.64 0.595 Proportion downward- Pilot Cue 8.43 0.006 swimming larvae Age 1.86 0.191 performing helices Cue x Age 1.07 0.375 Proportion of upward- Pilot Cue 1.56 0.223 swimming larvae Age 0.89 0.461 performing helices Cue x Age 1.90 0.156
Proportion larvae staing on bottom Fig. S1. Proportion of Crassostrea virginica larvae remaining on the bottom after swimming down as a function of larval age in the pilot experiment. Larvae were exposed to filtered seawater (no cue; light bars) or a chemical settlement cue (dark bars) in a pilot experiment. Error bars show standard error. 1.2 1 0.8 0.6 0.4 0.2 0 Hours post-competenc
Vertical swimming velcoit (cm/s) Vertical swimming velocit (cm/s) Fig. S2. Vertical swimming velocit of downward-swimming Crassostrea virginica larvae as a function of age in the pilot experiment. Larvae were exposed to filtered seawater (no cue; light bars) or a chemical settlement cue (dark bars) in a pilot experiment. Error bars show standard error. 0.0 Hours post-competenc -0.1-0.2-0.3-0.4-0.5 Downward-swimming larvae Fig. S3. Vertical swimming velocit of upward-swimming Crassostrea virginica larvae as a function of age in the pilot experiment. Larvae were exposed to filtered seawater (no cue; light bars) or a chemical settlement cue (dark bars) in a pilot experiment. Error bars show standard error. The missing error bar at t = 4 hours indicates onl one replicate had larvae swimming up off the bottom. Dissimilar letters indicate significant post hoc differences between ages, tested separatel for no-cue (a, b, c) and cue (, z) treatments. 0.15 0.10 0.05 b a a ab Upward-swimming larvae 0.00 Hours post-competenc
Gross to net distance ratio Gross to net distance ratio Fig. S4. Gross to net distance ratio (GNDR) of Crassostrea virginica larvae in the pilot experiment as a function of age for larvae swimming downward to the bottom of the flask following introduction (A) and larvae swimming back up off the bottom (B). Larvae were exposed to filtered seawater (no cue; light bars) or a chemical settlement cue (dark bars) in a pilot experiment. Error bars show standard error. The missing error bar at t = 4 hours indicates onl one replicate had larvae swimming up off the bottom. Dissimilar letters indicate significant post hoc differences between ages, tested separatel for no-cue (a, b, c) and cue (, z) treatments. 4 A 2 a ab b ab Downward-swimming larvae 0 Hours post-competenc 10 5 B Upward-swimming larvae 0 Hours post-competenc
Prop. larvae performing helices Prop. larvae performing helices Fig. S5. Proportion of Crassostrea virginica larvae in the pilot experiment swimming in helices as a function of age for larvae swimming downward to the bottom of the flask following introduction (A) and larvae swimming back up off the bottom (B). Larvae were exposed to filtered seawater (no cue, light bars) or a chemical settlement cue (dark bars) in a pilot experiment. Error bars show standard error. The missing error bar at t = 4 hours indicates onl one replicate had larvae swimming up off the bottom. 0.50 0.25 A Downward-swimming larvae 0.00 Hours post-competenc 1.0 B 0.5 Upward-swimming larvae 0.0 Hours post-competenc
Table S4. Non-parametric statistical results for heteroscedastic larval behavioral metrics. Mann-Whitne (M-W) and Kruskal-Wallis (K-W) tests show differences in the proportion of larvae remaining on the flask bottoms, downward swimming velocit of larvae, average gross:net distance ratio, and proportion swimming in helices. Factors are Cue (presence or absence of a settlement cue) and Age (larval age in hours post-competenc. Significant p- values (<0.05) shown in bold. Behavioral metric Experiment Test Factor χ 2 or U p Proportion larvae Pre and Post M-W Cue 133 < 0.001 remaining on bottom K-W Age 2.74 0.253 Velocit of downward- Time Series M-W Cue 663 0.453 swimming larvae K-W Age 1.14 0.887 GNDR of downward- Time Series M-W Cue 695 0.164 swimming larvae K-W Age 10.9 0.027 Pre and Post M-W Cue 297 0.007 K-W Age 3.31 0.191 GNDR of upward- Time Series M-W Cue 474 0.063 swimming larvae K-W Age 3.07 0.545 Proportion downward- Time Series K-W Age 8.01 0.091 swimming larvae Pre and Post M-W Cue 330 < 0.001 performing helices Proportion of upward- Time Series M-W Cue 514 0.388 swimming larvae performing helices K-W Age 2.94 0.568